title: High-resolution restoration of 3D structures from widefield images with extreme low signal-to-noise-ratio.
abstract: Four-dimensional fluorescence microscopy--which records 3D image information as a function of time--provides an unbiased way of tracking dynamic behavior of subcellular components in living samples and capturing key events in complex macromolecular processes. Unfortunately, the combination of phototoxicity and photobleaching can severely limit the density or duration of sampling, thereby limiting the biological information that can be obtained. Although widefield microscopy provides a very light-efficient way of imaging, obtaining high-quality reconstructions requires deconvolution to remove optical aberrations. Unfortunately, most deconvolution methods perform very poorly at low signal-to-noise ratios, thereby requiring moderate photon doses to obtain acceptable resolution. We present a unique deconvolution method that combines an entropy-based regularization function with kernels that can exploit general spatial characteristics of the fluorescence image to push the required dose to extreme low levels, resulting in an enabling technology for high-resolution in vivo biological imaging.
authors.affiliation: Department of Biochemistry and Biophysics and Keck Advanced Microscopy Center, University of California, San Francisco, CA 94158.
authors.forename: Muthuvel
authors.initials: M
authors.lastname: Arigovindan
authors.forename: Jennifer C
authors.initials: JC
authors.lastname: Fung
authors.forename: Daniel
authors.initials: D
authors.lastname: Elnatan
authors.forename: Vito
authors.initials: V
authors.lastname: Mennella
authors.forename: Yee-Hung Mark
authors.initials: YH
authors.lastname: Chan
authors.forename: Michael
authors.initials: M
authors.lastname: Pollard
authors.forename: Eric
authors.initials: E
authors.lastname: Branlund
authors.forename: John W
authors.initials: JW
authors.lastname: Sedat
authors.forename: David A
authors.initials: DA
authors.lastname: Agard
mesh_terms: D000465:Algorithms; D000818:Animals; D002460:Cell Line; D019277:Entropy; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D008958:Models, Molecular; D008962:Models, Theoretical; D009687:Nuclear Proteins / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D015203:Reproducibility of Results; D012441:Saccharomyces cerevisiae / Q000166:cytology / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000378:metabolism; D059629:Signal-To-Noise Ratio
chemical_list: D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; C090118:Zip1 protein, S cerevisiae
keywords: 4D microscopy; low dose microscopy; noise-suppressing regularization